Identification of Novel Trophoblast Invasion-Related Genes: Heme Oxygenase-1 Controls Motility via Peroxisome Proliferator-Activated Receptor γ
Identification of Novel Trophoblast Invasion-Related Genes: Heme Oxygenase-1 Controls Motility via Peroxisome Proliferator-Activated Receptor γ
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DOI：
10.1210/en.2008-0456
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发表时间：
2009-02-01
期刊：
影响因子：
4.8
通讯作者：
Knoefler, Martin
中科院分区：
文献类型：
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作者：
Bilban, Martin;Haslinger, Peter;Knoefler, Martin
细胞滋养层细胞（CTB）侵入子宫组织是胎盘发育所必需的。为了鉴定调节滋养层细胞侵袭的分子，分别从孕早期人类胎盘和绒毛外植体培养物中分离的纯化绒毛（CTB，侵袭性差）和绒毛外滋养层细胞（EVT）（侵袭性高）的mRNA特征，使用基因芯片分析进行比较，得到991个侵袭/迁移相关的转录本。参与生理和病理细胞侵袭的几个基因，包括A去整合素和金属蛋白酶-12、-19、-28，以及Spondin-2，在EVT中上调。通路预测分析确定了与侵袭性或非侵袭性滋养层表型相关的几个功能模块。选择在侵袭性mRNA库中下调的基因之一血红素加氧酶-1（HO-1）进行功能分析。实时PCR分析，蛋白质印迹法，和免疫荧光的第一个三个月胎盘和分化绒毛外植体培养证明下调HO-1在侵袭性EVT相比，CTBs。HO-1表达在功能丧失和功能获得细胞模型（分别为BeWo和HTR 8/SVneo）中的调节证明了HO-1表达与transwell和伤口愈合测定中的滋养层迁移呈反比关系。重要的是，HO-1表达导致核激素受体过氧化物酶体增殖物激活受体（PPAR）γ的蛋白水平和活性增加。药理学抑制的过氧化物酶体增殖物激活受体γ废除了HO-1对滋养层细胞迁移的抑制作用。总的来说，我们的研究结果表明，EVT和CTB的基因表达谱可以用来解开新的调节细胞侵袭。因此，我们确定HO-1作为一个负调节滋养层运动的作用，通过上调过氧化物酶体增殖物激活受体γ。（内分泌学150：1000-1013，2009）
Invasion of cytotrophoblasts (CTBs) into uterine tissues is essential for placental development. To identify molecules regulating trophoblast invasion, mRNA signatures of purified villous (CTB, poor invasiveness) and extravillous trophoblasts (EVTs) (high invasiveness) isolated from first trimester human placentae and villous explant cultures, respectively, were compared using GeneChip analyses yielding 991 invasion/migration-related transcripts. Several genes involved in physiological and pathological cell invasion, including A disintegrin and metalloprotease-12, -19, -28, as well as Spondin-2, were up-regulated in EVTs. Pathway prediction analyses identified several functional modules associated with either the invasive or noninvasive trophoblast phenotype. One of the genes that was down-regulated in the invasive mRNA pool, hemeoxygenase-1 (HO-1), was selected for functional analyses. Real-time PCR analyses, Western blotting, and immunofluorescence of first trimester placentae and differentiating villous explant cultures demonstrated down-regulation of HO-1 in invasive EVTs as compared with CTBs. Modulation of HO-1 expression in loss-of as well as gain-of function cell models (BeWo and HTR8/SVneo, respectively) demonstrated an inverse relationship of HO-1 expression with trophoblast migration in transwell and wound healing assays. Importantly, HO-1 expression led to an increase in protein levels and activity of the nuclear hormone receptor peroxisome proliferator activated receptor (PPAR) gamma. Pharmacological inhibition of PPAR gamma abrogated the inhibitory effects of HO-1 on trophoblast migration. Collectively, our results demonstrate that gene expression profiling of EVTs and CTBs can be used to unravel novel regulators of cell invasion. Accordingly, we identify HO-1 as a negative regulator of trophoblast motility acting via up-regulation of PPAR gamma. (Endocrinology 150: 1000-1013, 2009)